Novel IFN - Dependent Innate Antiviral Mechanisms

干扰素(IFN)在抵抗RNA和DNA病毒感染的先天防御中起着关键作用。 对这些关键细胞因子如何抑制病毒感染的详细了解可能会导致药理学 诱导对多种潜在危险制剂产生抗药性而无需诱导的方法 先前的抗原特异性免疫。鉴于大量潜在的生物武器，没有 疫苗的存在，这是一个重要的目标。干扰素通过诱导或抑制干扰蛋白发挥作用 病毒复制或改变免疫细胞的功能。在这笔赠款中，我们将重点研究受干扰素调控的基因 抑制病毒复制。有数百个受干扰素调控的基因，其中只有几个的功能是 理解得很详细。筛选多种干扰素调控基因以寻找具有特异性抗病毒作用的基因 功能是一项艰巨的任务。我们的初步数据和其他人的工作表明，这一点至关重要 干扰素诱导的抗病毒分子仍未确定。我们已经成功地开发出了 发现这些重要的抗病毒分子。 基于这些考虑，我们建议对小说的作用机制进行识别和分析 干扰素诱导的抗病毒蛋白。我们将使用两个筛选抗病毒蛋白：(I)表达克隆 阻止病毒复制的蛋白质，以及(Ii)使用与Sindbis载体相连的基因芯片分析 互补实验用于鉴定体内具有抗病毒功能的蛋白质。我们已经成功地利用了这一点 鉴定干扰素诱导的抗病毒蛋白的第二种方法，泛素样蛋白ISG15。我们有 开始分析ISG15在抵抗包括A类和B类病毒在内的几种病毒中的作用 我们将详细分析ISG 15在受感染细胞中的作用(目标1)，并继续使用我们的屏幕 已经建立以鉴定额外的干扰素诱导的抗病毒分子(目标2)。 目的1.确定ISG15抗病毒作用的分子机制(S)。 目的2.筛选新的抗病毒候选蛋白，并证明其生理意义。

Interferons (IFNs) play a critical role in innate defense against infection with RNA and DNA viruses. A detailed understanding of how these critical cytokines inhibit viral infection might lead to pharmacologic approaches to induce resistance to multiple potentially dangerous agents without the necessity of inducing prior antigen-specific immunity. Given the large number of potential biological weapons for which no vaccine exists, this is an important goal. IFNs work via induction or repression of proteins that interfere with viral replication or alter the function of immune cells. In this grant we will focus on IFN regulated genes that inhibit viral replication. There are hundreds of IFN regulated genes, the function of only a few of which is understood in detail. Sorting through the many IFN regulated genes to find those with specific antiviral functions is a daunting task. Our preliminary data, and work from others, suggests that critically important IFN inducible antiviral molecules remain unidentified. We have succeeded in developing methods for discovering these important antiviral molecules. Based on these considerations, we propose to identify and analyze the mechanisms of action of novel IFN induced antiviral proteins. We will use two screens for antiviral proteins: (i) expression cloning of proteins that block viral replication, and (ii) use of gene chip analysis coupled to a Sindbis vector based complementation assay to identify proteins with antiviral function in vivo. We have successfully used this second approach to identify an IFN inducible antiviral protein, the ubiquitin-like protein ISG15. We have begun to analyze the role of ISG15 in resistance to several viruses including viruses in Class A and B. In this proposal we will analyze ISG 15 action in infected cells in detail (Aim 1), and continue to use the screens we have set up to identify additional novel IFN induced antiviral molecules (Aim 2). Aim 1. Determine the molecular mechanism(s) of the antiviral effect of ISG15. Aim 2. Identify candidate novel antiviral proteins and prove their physiologic relevance.